Fig. 4

Inhibition of NR5A2 specifically eliminates pancreatic cancer stem cells. A Flow cytometry analysis of CD133⁺ CSCs following 72 h of treatment with siNR5A2 variants #1 and #2; 'src' indicates siScramble. Representative flow cytometry dot plots are displayed. B Quantification of n = 3 biological replicates. C Flow cytometry analysis of CD133⁺ CSCs following 72 h of treatment with graded doses of Cpd3. Representative data are depicted. D Quantification of CD133⁺ CSCs in n = 3 biological replicates. E Percentage of apoptotic Annexin V positive cells among CD133^– differentiated cancer cells (grey) and CD133⁺ CSC (blue) following treatment with siNR5A2 variants #1 and #2, and F the percentage among CD133^– differentiated cancer cells (grey) and CD133⁺ CSC (blue) following treatment with 20 and 40 µM Cpd3 in n = 3 biological replicates. G Immunofluorescence for Pan-cytokeratin (green) following 48 h of treatment with 80 µM Cpd3. Nuclei were stained with DAPI (red). H In vivo tumorigenicity of decreasing numbers of highly enriched CD133⁺ FLUO⁺ CSCs following pharmacological or genetic targeting of NR5A2. I Flow cytometry for CD133⁺ CD44⁺ and CD133⁺ CXCR4⁺ CSCs in harvested tumors. In panels A to F, data are presented as mean ± SD and statistically analyzed using two-tailed Mann–Whitney tests to compare two groups (n = 3 biological replicates). In panel I, data are presented as floating bars, and statistically analyzed using two-tailed Mann–Whitney tests to compare two groups (n = 3–6 tumors). Asterisks indicate significance at the indicated levels: * p < 0.05. Please also see Supplementary Fig. 4